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Biomedical subjects

R K Singh

Publications and source records attributed to R K Singh.

At least 73 records · Page 4Linked to original sources
PubMed2002

Polymerase chain reaction amplification of 16S-23S spacer region for rapid identification of Salmonella serovars.

Polymerase chain reaction (PCR) was used to amplify the spacer regions between the 16S and 23S genes of rRNA genetic loci of Salmonella serovars for their rapid identification. These genetic loci revealed a significant level of polymorphism in length across the species/serovar lines. When the 16S-23S spacer region amplification products were subjected to agarose electrophoresis, the patterns observed could be used to distinguish all the serovars of Salmonella tested. Unique elements obtained in amplification products were mostly clustered at serovar level, although certain genus-specific patterns were also observed. On the basis of the results obtained, the amplification of 16S-23S ribosomal spacer region could suitably be used in a PCR-based identification method for Salmonella serovars.

Animals↗
PubMed2002

Sarcoglycanopathies: a report of 25 cases.

Twenty five patients with sarcoglycanopathies were studied prospectively. 21 of them had mild phenotype. Muscle involvement was more pronounced in adductor and flexor groups of muscles of the limbs, hip adductor muscles being the weakest. The selective and differential weakness between weak hip adductors and stronger hip abductors resulted in the hip abduction sign in 64% of cases. Distal muscle involvement in lower limbs was seen in 92% of cases, but was mild and late in the course of the disease. 44% patients had winging of scapulae. Immunocytochemistry showed multiple sarcoglycan deficiencies in 84% patients. Primary beta and delta sarcoglycanopathy was seen in the remaining 16% cases. Secondary dystrophin reduction was seen in 44% patients and correlated with beta sarcoglycan deficiency but not with functional disability.

Adolescent↗
PubMed2002

Acute febrile neutrophilic dermatosis following tuberculous infection.

A case of acute febrile neutrophilic dermatosis (Sweet's syndrome) in association with tuberculosis has been reported. However association of the two diseases is not common. It appears as if Mycobacterium tuberculosis present somewhere in the body can induce cutaneous reactions as is the case in erythema nodosum and erythema induratum.

Acute Disease↗
PubMed2001

Characterization of wet and dry deposition in the downwind of industrial sources in a dry tropical area.

An atmospheric deposition study was conducted in the downwind of Shaktinagar Thermal Power Plant (STPP), Renusagar Thermal Power Plant (RTPP), and Anpara Thermal Power Plant (ATPP), at Singrauli region, Uttar Pradesh (UP), India to characterize dry and wet deposition in relation to different pollution loading. During the study period, dry and wet depositions and levels of gaseous pollutants (SO2 and NO2) were estimated across the sites. Dry deposition was collected on a monthly basis and wet deposition on an event basis. Depositions were analyzed for pH, nitrate (NO3-), ammonium (NH4+), and sulphate (SO4(2-)) contents. Dry deposition rate both collected as clearfall and throughfall varied between 0.15 to 2.28 and 0.33 to 3.48 g m(-2) day(-1), respectively, at control and maximally polluted sites. The pH of dry deposition varied from 5.81 to 6.89 during winter and 6.09 to 7.02 during summer across the sites. During the rainy season, the mean pH of clear wet deposition varied from 6.56 to 7.04 and throughfall varied from 6.81 to 7.22. The concentrations of NO2 and SO2 pollutants were highest during the winter season. Mean SO2 concentrations varied from 18 to 75 g m(-3) at control and differently polluted sites during the winter season. The variation in NO2 concentrations did not show a pattern similar to that of SO2. The highest NO2 concentration during the winter season was 50 g m(-3), observed near RTPP. NO2 concentration did not show much variation among different sites, suggesting that the sources of NO2 emission are evenly distributed along the sites. The concentrations of NH4+, NO3-, and SO4(2-) ions in dry deposition were found to be higher in summer as compared to the winter season. In dry deposition (clearfall) the concentrations of NH4+, NO3-, and SO4(2-) varied from 0.13 to 1.0, 0.81 to 1.95, and 0.82 to 3.27 mg l(-1), respectively, during winter. In wet deposition (clearfall), the above varied from 0.14 to 0.74, 0.81 to 1.82, and 0.67 to 2.70 mg l(-1), respectively. The study clearly showed that both dry and wet depositions varied between the sites and season, suggesting significant impact of industrial activities in modifying the atmospheric input. The nonacidic deposition suggests that there is no threat of acidification of the receiving ecosystem at present.

Air Pollutants↗
PubMed2001

The complete genome of the crenarchaeon Sulfolobus solfataricus P2.

The genome of the crenarchaeon Sulfolobus solfataricus P2 contains 2,992,245 bp on a single chromosome and encodes 2,977 proteins and many RNAs. One-third of the encoded proteins have no detectable homologs in other sequenced genomes. Moreover, 40% appear to be archaeal-specific, and only 12% and 2.3% are shared exclusively with bacteria and eukarya, respectively. The genome shows a high level of plasticity with 200 diverse insertion sequence elements, many putative nonautonomous mobile elements, and evidence of integrase-mediated insertion events. There are also long clusters of regularly spaced tandem repeats. Different transfer systems are used for the uptake of inorganic and organic solutes, and a wealth of intracellular and extracellular proteases, sugar, and sulfur metabolizing enzymes are encoded, as well as enzymes of the central metabolic pathways and motility proteins. The major metabolic electron carrier is not NADH as in bacteria and eukarya but probably ferredoxin. The essential components required for DNA replication, DNA repair and recombination, the cell cycle, transcriptional initiation and translation, but not DNA folding, show a strong eukaryal character with many archaeal-specific features. The results illustrate major differences between crenarchaea and euryarchaea, especially for their DNA replication mechanism and cell cycle processes and their translational apparatus.

Cell Cycle Proteins↗
PubMed2001

Kawasaki disease.

Kawasaki disease (KD) is a systemic necrotizing vasculitis affecting medium and small sized arteries. The diagnosis is based entirely on recognition of a typical sequence of clinical features. Detection of any one clinical feature does not have any diagnostic significance. We report an uncommon case of Kawasaki disease in 10 months old male child with the analysis of its natural history, etiopathology, treatment and prognosis of the disease.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2001

GB syndrome with herpes simplex infection.

G.B. Syndrome is an acute flaccid lower motor neuron (LMN) paralysis. The diagnosis is based on clinical presentation, course of illness with supportive investigations. This article reports an uncommon case of GB Syndrome caused by HSV infection in a 2 yr-9 mth-old child and have analysed the natural history, etiopathology, treatment and prognosis of the disease.

Child, Preschool↗
PubMed2001

Flt3 ligand and granulocyte-macrophage colony-stimulating factor preferentially expand and stimulate different dendritic and T-cell subsets.

OBJECTIVE: Mechanisms of T-cell stimulation by Flt3 ligand (Flt3L) and granulocyte-macrophage colony-stimulating factor (GM-CSF) remain unclear. Herein, we compared the effects of Flt3L and GM-CSF on the expansion of dendritic cells (DC) and T-cell subsets and cytokine expression. METHODS: Naïve and effector/memory T cells were analyzed by flow cytometry (FC). CD4(+) and CD8(+) T cells and CD11c(+)CD11b(dull/-)(DC1) and CD11c(+)CD11b(+) (DC2) subsets were isolated and the frequency of IFN-gamma-, IL-12- (type 1) and IL-4-, IL-10 (type 2)-producing cells and cytokine mRNA expression evaluated. RESULTS: Flt3L expanded both DC1 and DC2 subsets with a significantly higher percentage and number of DC1 than DC2, while GM-CSF preferentially expanded the DC2 subset. Isolated DC1 from Flt3L-injected mice had significantly higher levels of IL-12 (p40) than IL-10, while the converse occurred with DC2. The numbers of naïve and memory T cells were elevated in mice that received Flt3L or GM-CSF. However, the number of memory CD4(+) and CD8(+) T cells was significantly increased in Flt3L as compared to GM-CSF cohorts. While GM-CSF increased the frequency of both type 1 and type 2 cytokine-producing cells, Flt3L significantly augmented the frequency of type 1 T cells. CONCLUSIONS: In contrast to GM-CSF, Flt3L preferentially induces the expansion of type 1 T cells. The mechanism of Flt3L-induced T-cell stimulation is associated with the expansion of the IL-12 (p40)-producing DC1 and memory T cells.

Animals↗
PubMed2001

Anti-inflammatory effect of (+)-pinitol.

In the carrageenin-induced paw oedema in rats, (+)-pinitol (2.5-10 mg/kg, i.p.), isolated from Abies pindrow leaves, showed a significant anti-inflammatory effect, the highest dose being comparable to phenylbutazone (100 mg/kg, i.p.).

Animals↗
PubMed2001

Activation-induced T cell apoptosis by monocytes from stem cell products.

We recently found that mobilized peripheral blood stem cell (PSC) products (from both cancer patients and normal donors) contain high levels of CD14+ monocytes, which can inhibit the proliferation of allogeneic and autologous T cells. We found in our studies that using CD14+ monocytes from mobilized PSC products (from normal and cancer patient donors), normal apheresis products or normal peripheral blood (PB) can affect lymphocyte function and apoptosis-dependent T cell activation. However, it appears that the apoptosis is dependent on the frequency of monocytes, which is increased by both mobilization and apheresis. Both phytohemagglutinin (PHA)- and interleukin (IL)-2-induced proliferation of steady-state peripheral blood mononuclear cells (PBMC) were markedly inhibited by co-culture with irradiated CD14+ monocytes, although inhibition was significantly greater with PHA than with IL-2 stimulation. IL-2 (predominately CD56+ NK cells) or anti-CD3 monoclonal antibody (mAb) and IL-2-expanded lymphocytes (activated T cells) were inhibited by PSC monocytes to a significantly greater level as compared to steady-state lymphocytes. Indeed, no inhibition of T cell proliferation was observed when lymphocytes were co-cultured in the absence of mitogenic or IL-2 stimulation. In contrast, an increased proliferation was observed in co-cultures of CD14+ monocytes and steady-state or activated lymphocytes without mitogenic stimulation. Cell cycle analysis by flow cytometry revealed a significant increase in hypodiploid DNA, in a time-dependent manner, following co-culture of monocytes and PBMC in PHA, suggesting that T cell apoptosis occurred during PHA-induced activation. These results demonstrate that PSC-derived monocytes inhibit T cell proliferation by inducing the apoptosis of activated T cells and NK cells, but not steady-state cells. This suggests a potential role for monocytes in the induction of peripheral tolerance following stem cell transplantation.

Antibodies, Monoclonal↗
PubMed2001

Preliminary report on a single-tube, non-interrupted reverse transcription-polymerase chain reaction for the detection of rabies virus in brain tissue.

A simple method for the rapid detection of rabies virus was developed employing a single-tube reverse transcription polymerase chain reaction (RT-PCR). The method utilized a single buffer system for both RT and PCR and was performed without interruption as a single thermal cycling programme. Two primer sets within the genes coding for rabies nucleoprotein and glycoprotein were used to amplify a 533 bp and a 406 bp amplicon, respectively. The amplified products were detected with a challenge virus strain (CVS) of rabies. There was no amplified product with uninfected mouse or dog brain. The method was applied to detect rabies virus in 10 mouse inoculation test (MIT)-positive and three MIT-negative brain tissue samples. The amplified product was found only in the MIT-positive samples. The amplified product was confirmed by restriction endonuclease analysis using Hinf1. The results from RT-PCR correlated well with the results from MIT. This indicates that the single-tube RT-PCR may be a useful method for detecting rabies virus in brain tissue samples from suspected cases of rabies.

Animals↗
PubMed2001

Enhancement of enzyme activity through three-phase partitioning: crystal structure of a modified serine proteinase at 1.5 A resolution.

Three-phase partitioning is fast developing as a novel bioseparation strategy with a wide range of applications including enzyme stability and enhancement of its catalytic activity. Despite all this, the enzyme behaviour in this process still remains unknown. A serine proteinase, proteinase K, was subjected to three-phase partitioning (TPP). A 3 ml volume of proteinase K solution (3 mg/ml in 0.05 M acetate buffer, pH 6.0) was brought to 30% (w/v) ammonium sulphate saturation by addition of saturated ammonium sulphate. tert-Butanol (6 ml) was added to this solution and the mixture was incubated at 25 degrees C for 1 h. The precipitated protein in the mid-layer was dissolved in 3 ml of 0.05 M acetate buffer, pH 6.0. The specific activity of the processed enzyme was estimated and was found to be 210% of the original enzyme activity. In order to understand the basis of this remarkable enhancement of the enzyme activity, the structure of the TPP-treated enzyme was determined by X-ray diffraction at 1.5 A resolution. The overall structure of the TPP-treated enzyme is similar to the original structure in an aqueous environment. The hydrogen bonding system of the catalytic triad is intact. However, the water structure in the substrate binding site has undergone a rearrangement as some of the water molecules are either displaced or completely absent. Two acetate ions were identified in the structure. One is located in the active site and seems to mimic the role of water in the enzyme activity and stability. The other is located at the surface of the molecule and is involved in stabilizing the local structure of the enzyme. The most striking observation in respect of the present structure pertains to a relatively higher overall temperature factor (B = 19.7 A(2)) than the value of 9.3 A(2) in the original enzyme. As a result of a higher B-factor, a number of residues, particularly their side chains, were found to adopt more than one conformation. It appears that the protein exists in an excited state which might be helping the enzyme to function more rapidly than the original enzyme in aqueous media. Summarily, the basis of increased enzymatic activity could be attributed to (i) the presence of an acetate ion at the active site and (ii) its excited state as reflected by an overall higher B-factor.

Crystallography, X-Ray↗